机构:[1]Cell Therapy Center, Beijing Municipal Geriatric Medical Research Center, Xuanwu Hospital Capital Medical University首都医科大学宣武医院[2]National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education,Xuanwu Hospital Capital Medical University.科技平台神经变性病教育部重点实验室首都医科大学宣武医院[3]The Department of Neurology, Xuanwu Hospital Capital Medical University.神经科系统神经内科首都医科大学宣武医院
The integration of designer receptors exclusively activated by designer drugs (DREADD) with stem cell-based therapies presents an advanced strategy for precise neuronal modulation. Here, we utilized CRISPR-engineered human reprogrammed stem cells expressing excitatory (hM3Dq) or inhibitory (hM4Di) DREADD receptors to evaluate the functional integration and modulation of transplanted dopaminergic precursors in a murine model of Parkinson's disease (PD). Key steps included generating non-fusion DREADD-expressing stem cell lines, differentiating them into midbrain dopaminergic precursors, and transplanting these cells into the striatum of 6-hydroxydopamine (6-OHDA)-lesioned mice. We conducted behavioral assessments and electrophysiological recordings to analyze the effects of the transplanted cells. Behavioral tests, such as the cylinder test, demonstrated significant modulation of motor function following clozapine-N-oxide (CNO) administration. Specifically, activation of hM4Di reduced contralateral forelimb movement, whereas activation of hM3Dq was associated with enhanced motor behavior. Electrophysiological recordings revealed distinct synaptic responses. hM4Di activation increased interevent intervals and decreased peak amplitudes of spontaneous excitatory postsynaptic currents (sEPSCs), whereas hM3Dq activation decreased interevent intervals and increased peak amplitudes, reflecting enhanced excitatory signaling. In summary, the integration of behavioral and electrophysiological assessments validates the precise functional incorporation of engineered chemically reprogrammed stem cells into host neural circuits.
基金:
This work was supported by the Beijing Natural Science
Foundation (7242068), National Natural Science Foundation
of China (82171250), Beijing Municipal Health Commission
Fund (PXM2020_026283_000005) and The Project for
Technology Development of Beijing-affiliated Medical
Research Institutes (11000023T000002036310).
语种:
外文
PubmedID:
中科院(CAS)分区:
出版当年[2025]版:
大类|4 区综合性期刊
小类|4 区综合性期刊
最新[2025]版:
大类|4 区综合性期刊
小类|4 区综合性期刊
第一作者:
第一作者机构:[1]Cell Therapy Center, Beijing Municipal Geriatric Medical Research Center, Xuanwu Hospital Capital Medical University[2]National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education,Xuanwu Hospital Capital Medical University.
通讯作者:
通讯机构:[1]Cell Therapy Center, Beijing Municipal Geriatric Medical Research Center, Xuanwu Hospital Capital Medical University[2]National Clinical Research Center for Geriatric Diseases, and Key Laboratory of Neurodegenerative Diseases, Ministry of Education,Xuanwu Hospital Capital Medical University.
推荐引用方式(GB/T 7714):
Han Deqiang,Wang Xueyao,Pan Na,et al.Chemogenetic Regulation in Reprogrammed Stem Cell-derived Precursor Cells in Treating Neurodegenerative Diseases[J].Journal Of Visualized Experiments : Jove.2025,(219):doi:10.3791/68032.
APA:
Han Deqiang,Wang Xueyao,Pan Na,Jing Shuili&Chen Zhiguo.(2025).Chemogenetic Regulation in Reprogrammed Stem Cell-derived Precursor Cells in Treating Neurodegenerative Diseases.Journal Of Visualized Experiments : Jove,,(219)
MLA:
Han Deqiang,et al."Chemogenetic Regulation in Reprogrammed Stem Cell-derived Precursor Cells in Treating Neurodegenerative Diseases".Journal Of Visualized Experiments : Jove ..219(2025)